Information on EC 1.14.13.78 - ent-kaurene oxidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.78
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RECOMMENDED NAME
GeneOntology No.
ent-kaurene oxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ent-kaur-16-en-19-al + NADPH + O2 = ent-kaur-16-en-19-oate + NADP+ + H2O
show the reaction diagram
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-
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ent-kaur-16-en-19-ol + NADPH + H+ + O2 = ent-kaur-16-en-19-al + NADP+ + 2 H2O
show the reaction diagram
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-
-
-
ent-kaur-16-ene + NADPH + H+ + O2 = ent-kaur-16-en-19-ol + NADP+ + H2O
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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-
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Diterpenoid biosynthesis
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Metabolic pathways
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
ent-kaur-16-ene,NADPH:oxygen oxidoreductase (hydroxylating)
Requires cytochrome P-450. Catalyses three sucessive oxidations of the 4-methyl group of ent-kaurene giving kaurenoic acid.
CAS REGISTRY NUMBER
COMMENTARY hide
149565-67-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain SG78
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-
Manually annotated by BRENDA team
i.e. Gibberella fujikuroi or G. intermedia, MP-C or MP-D, strains IMI58289 and D02945, gene P450-4
SwissProt
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
wild cucumber
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Manually annotated by BRENDA team
bitter melon
UniProt
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(-)-kaurene + 3 NADPH + 3 H+ + 3 O2
(-)-kaurenoic acid + 3 NADP+ + 4 H2O
show the reaction diagram
ent-kaur-16-en-19-al + NADPH + O2
ent-kaur-16-en-19-oate + NADP+ + H2O
show the reaction diagram
ent-kaur-16-en-19-ol + NADPH + H+ + O2
ent-kaur-16-en-19-al + NADP+ + 2 H2O
show the reaction diagram
ent-kaur-16-ene + 3 NADPH + 3 H+ + 3 O2
ent-kaur-16-en-19-oate + 3 NADP+ + 4 H2O
show the reaction diagram
ent-kaur-16-ene + NADPH + H+ + O2
ent-kaur-16-en-19-ol + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene + NADPH + H+ + O2
ent-kaur16-en-19-ol + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene + NADPH + O2
ent-kaur-16-en-19-ol + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene-19-al + NADPH + H+ + O2
ent-kaur-16-en-19-oate + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene-19-al + NADPH + O2
ent-kaur-16-en-19-oic acid + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene-19-ol + NADPH + H+ + O2
ent-kaur-16-en-19-al + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene-19-ol + NADPH + O2
ent-kaur-16-en-19-al + NADP+ + H2O
show the reaction diagram
ent-kaur16-en-19-al + NADPH + H+ + O2
ent-kaur16-en-19-oate + NADP+ + H2O
show the reaction diagram
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three step oxidation process via ent-kaurenol and ent-kaurenal, in co-operation with membrane-associated cytochrome P450 monooxygenase
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-
?
ent-kaur16-en-19-ol + NADPH + H+ + O2
ent-kaur16-en-19-al + NADP+ + H2O
show the reaction diagram
ent-kaurene + NADPH + H+ + O2
(-)-kaurenoic acid + NADP+ + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(-)-kaurene + 3 NADPH + 3 H+ + 3 O2
(-)-kaurenoic acid + 3 NADP+ + 4 H2O
show the reaction diagram
ent-kaur-16-en-19-al + NADPH + O2
ent-kaur-16-en-19-oate + NADP+ + H2O
show the reaction diagram
ent-kaur-16-en-19-ol + NADPH + H+ + O2
ent-kaur-16-en-19-al + NADP+ + 2 H2O
show the reaction diagram
ent-kaur-16-ene + NADPH + H+ + O2
ent-kaur-16-en-19-ol + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene + NADPH + H+ + O2
ent-kaur16-en-19-ol + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
ent-kaur-16-ene + NADPH + O2
ent-kaur-16-en-19-ol + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene-19-al + NADPH + H+ + O2
ent-kaur-16-en-19-oate + NADP+ + H2O
show the reaction diagram
O94142
involved in gibberilin biosynthesis pathway
-
?
ent-kaur-16-ene-19-al + NADPH + O2
ent-kaur-16-en-19-oic acid + NADP+ + H2O
show the reaction diagram
ent-kaur-16-ene-19-ol + NADPH + H+ + O2
ent-kaur-16-en-19-al + NADP+ + H2O
show the reaction diagram
O94142
involved in gibberilin biosynthesis pathway
-
?
ent-kaur-16-ene-19-ol + NADPH + O2
ent-kaur-16-en-19-al + NADP+ + H2O
show the reaction diagram
ent-kaur16-en-19-al + NADPH + H+ + O2
ent-kaur16-en-19-oate + NADP+ + H2O
show the reaction diagram
-
three step oxidation process via ent-kaurenol and ent-kaurenal, in co-operation with membrane-associated cytochrome P450 monooxygenase
-
-
?
ent-kaur16-en-19-ol + NADPH + H+ + O2
ent-kaur16-en-19-al + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
ent-kaurene + NADPH + H+ + O2
(-)-kaurenoic acid + NADP+ + H2O
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FAD
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stimulates activity
NADPH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1E)-1-[4-(4-butyl-1H-1,2,3-triazol-1-yl)phenyl]-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)pent-1-en-3-ol
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(1E,3S)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)pent-1-en-3-ol
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i.e. uniconazole
(2R,3R)-paclobutrazol
(2S,3S)-paclobutrazol
(E)-2-(2-((1-(4-(3-hydroxy-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)pent-1-en-1-yl)phenyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethyl 4-methylbenzenesulfonate
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i.e. abscinazole-E1, orUT1-E2Ts, or Abz-E1, a specific potent inhibitor of ABA 8'-hydroxylase, EC 1.14.13.93, but a weak inhibitor of CYP701A, both in vitro and in vivo
abscinazole-F1
CO
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90% inhibition
DSI-505ME
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methyl (2E)-3-[1-[(1E)-1-(4-chlorophenyl)-3-hydroxy-4,4-dimethylpent-1-en-2-yl]-1H-imidazol-5-yl]prop-2-enoate
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paclobutrazol
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paclobutrazol racemate
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uniconazole
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome P450 monooxygenase
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necessary for ent-kaurenoate production by ent-kaurene oxidase
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0063
ent-kaur-16-ene
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00006 - 0.0015
(2R,3R)-paclobutrazol
0.00002 - 0.00007
(2S,3S)-paclobutrazol
0.00097
(E)-2-(2-((1-(4-(3-hydroxy-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)pent-1-en-1-yl)phenyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethyl 4-methylbenzenesulfonate
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pH 7.25, 30°C, recombinant enzyme
0.00002
paclobutrazol racemate
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.25
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assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.4
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
the enzyme expression in pistillate flowers is significantly higher than in staminate flowers and neutral flowers
Manually annotated by BRENDA team
transcript detection
Manually annotated by BRENDA team
additional information
the expression level is especially abundant in root and stem apexes
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
enzyme is targeted to the outer chloroplast envelope
Manually annotated by BRENDA team
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membrane associated
Manually annotated by BRENDA team
additional information
-
subcellular compartmentation, overview
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Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000 - 55000
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gel filtration
70000
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Western blot analysis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
20% glycerol reduces the degradation to P-420
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
25% polyethylene 8000, anion exchange, gel filtration
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recombinant CYP701A partially from insect Sf9 cells in microsomes
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recombinant CYP701A3 partially in microsomal membranes from Escherichia coli
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of cDNA
CYP701A, expression in Spodoptera frugiperda Sf9 cells using the baculovirus transfection method
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CYP701A3, functional expression in Escherichia coli using a fully codon-optimized construct, along with additional N-terminal deletion and modification
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DNA and amino acid sequence determimnation and analysis, expression in Escherichia coli, Saccharomyces cerevisiae, and in transgenic Arabidopsis thaliana leaves as GFP-tagged protein via Agrobacterium tumefaciens transfection, expression of the GFP-tagged enzyme in onion bulb epidermis or tobacco seedling leaves using particle bombardment method
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DNA and amino acid sequence determination and analysis, sequence comparisons; KOS2, DNA and amino acid sequence determination and analysis, sequence comparisons; KOS4, DNA and amino acid sequence determination and analysis, sequence comparisons
Q68YV7, Q68YV9, Q6GZ42
expressed in Escherichia coli BL21 cells
expressed in Escherichia coli BL21(DE3) cells
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expressed in Pichia pastoris
expression in Saccharomyces cerevisiae
expression of cDNA in Saccharomyces cerevisiae
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gene LsKO1, DNA and amino acid sequence determination and analysis, quantitative PCR expression analysis, expression in Pichia pastoris; gene LsKO2, DNA and amino acid sequence determination and analysis, quantitative PCR expression analysis, expression in Pichia pastoris
gene P450-4 from strain D02945, DNA and amino acid sequence determination and analysis, organization of the GA gene cluster, expression in Escherichia coli and in strain D02945, screening for ectopic integrations, P450-4 promoter studies using the GUS reporter system, expression analysis
KOS3, DNA and amino acid sequence determination and analysis, sequence comparisons
Q68YV8
overexpression in Arabidopsis thaliana
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PCR-amplification of cDNA, cloning into pPICZ yeast expression vector, expression in Pichia pastoris strain X-33 together with fungal P450 reductase (CPR)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
molecular biology
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the fungal cytochrome P450 monooxygenase isolated from from Phaeoshaeria sp., strain L487 in the Pichia pastoris expression system might become a tool for functional assays of a variety of rice cytochrome P450 monooxygenases involved in the biosynthesis of secondary metabolites